Development of Off-the-shelf Completely Biological Small-Diameter Blood Vessel with Human Stem Cells Project Summary Vascular grafts are in great demand due to the high occurrence of cardiovascular diseases. Despite the successful replacement of large-diameter blood vessels, small diameter blood vessels (such as coronary arteries) lack suitable replacement materials. Presently, no biomaterial or cell-based tissue-engineered vascular grafts (TEVG) can meet the urgent needs of patients for coronary artery substitutes. Although there has been great progress in developing cell sheet technology for tissue engineered vascular grafts, there is a striking deficit in our understanding of how the human cell donor characteristics influence the engineered material. Our main hypothesis is that a standardized extracellular matrix (ECM) tube with a three- dimensional (3D) interwoven nanofibrous organization, mimicking the cellular organization of natural blood vessels, will enable the reproducible production of a completely biological, mechanically strong, and anti- thrombogenic TEVG within a short time, upon the inclusion of an anti-thrombogenic cell type in the lumen of ECM tube. The objective of this project is to engineer and optimize a mechanically strong ECM tube using a highly aligned nanofibrous scaffold and then combine it with an antithrombogenic cell type to biofabricate a mechanically strong off-the-shelf or personalized small-diameter TEVG within a short time. Aim I: Develop a robust scheme for manufacturing an ECM tube with optimal 3D interwoven nanofibrous organization. Aim II: Create a completely biological TEVG by including hMSCs in the ECM tube. Aim III: Evaluate the tissue regeneration and long-term patency of the TEVG, in vivo. Aim IV Continue to Engage undergraduate students in stem cell engineering, tissue engineering, and biomaterials research at Michigan Tech.